阿霉素诱导人卵巢癌细胞耐药株的建立及多药耐药表型测定

Multidrug resistance phenotypic determination of ovarian cancer cell line induced by adriamycin

[目的]探讨卵巢癌细胞多药耐药的发生机制,寻找克服及逆转卵巢癌多药耐药的方法。[方法]采用阿霉素较大剂量间歇诱导和浓度梯度递增培养法,建立人卵巢癌耐药细胞株(SK-OV-3/adr)。运用MTT法检测药物敏感性和细胞生长规律变化,流式细胞仪检测细胞周期分布,荧光显微镜观察耐药株细胞内药物含量变化。[结果]建立的SK-OV-3/adr对阿霉素、5-氟尿嘧啶、长春新碱、紫杉醇、足叶乙甙有明显的交叉耐药,耐药指数分别为5.6(296.7/53.4 ng.mL-1)、6.3(12008.4/1921.6 ng.mL-1)、4.6(1176.2/254.3 ng.mL-1)、12.5(4269.6/342.0ng.mL-1)、>10.9(>20000/1830.1 ng.mL-1)。与卵巢癌细胞敏感株(SK-OV-3)相比,半数细胞抑制剂量IC50明显增加,差异有统计学意义(P<0.01)。给予P-糖蛋白抑制剂(盐酸维拉帕米)时,交叉耐药性减弱。与SK-OV-3相比,SK-OV-3/adr细胞内阿霉素荧光强度明显变弱。[结论]建立了具有多药耐药表型的SK-OV-3/adr细胞株。

[ Objective ] To explore the muhidrug resistance mechanism in ovarian carcinoma. [ Method ] The human ovarian carcinoma cell line of resistant to adriamycin （SK- OV -3/adr） was established through progressively increased concentration and intermittent administration of high dose of adriamycin. Drug sensitivity and cell growth change were examined by MTT assay. Cell cycle distribution was investigated by flow cytometry. The drug level of the resistant line was observed by. [ Results ] The cell line SK - OV - 3/adr was established, which showed an obvious cross - resistance to adriamycin, 5 -fluorouracil ,vincristine and taxol Vpl6 steadily. The index of muhidrug resistance were 5.6 （296.7/53.4 ng·mL^-1）, 6.3 （ 12008.4/1921.6 ng·mL^-1）, 4.6 （ 1176.2/254.3 ng·mL^-1 ）, 12.5 （4269.6/342.0 ng·mL^-1 ）, 〉 10.9 （ 〉 20000/1830.1 ng·mL^-1 ） respectively. IC50 of SK - OV - 3/adr increased significantly compared with that of the sensitive strain of ovarian carcinoma （ SK - OV - 3 ） , P 〈 0.01. The effect decreased when P - glycoprotein inhibitor （ verapamil hydrochloride） was added. The adriamycin fluorescence intensity in SK - OV - 3/adr cells was significantly weaker than that in SK - OV - 3 cells. [ Conclusion ] The multidrug resistant cell line of SK - OV - 3/adr was successfully established.